Secretion of hirudin derivatives
A process for obtaining hirudin derivatives from E. coli secretor mutants which entails:(1) construction of a recombinant vector on which there is located the gene coding for a hirudin derivative directly downstream of a DNA section which codes for a bacterial signal peptide;(2) transformation of an E. coli secretor mutant with the recombinant vector constructed in step (1);(3) cultivation of the transformed cells in a medium; and(4) obtaining the hirudin derivative from the medium; anda recombinant vector which contains one or more copies of a gene construct which codes for a protein consisting of a bacterial signal peptide and of a hirudin derivative, and a hirudin derivative with the N-terminal amino-acid sequence A-(SEQ ID NO: 5) in which A represents Ala, Gln, His, Phe, Tyr, Glu, Ser, Asp or Asn.
Latest Consortium fur elektrochemische Industrie GmbH Patents:
- Method for the production of organosiloxanes modified by a phosponic acid ester
- Method for the production of isocyanatoorganosilanes
- Rapid-cure, one-component mixtures, which contain alkoxysilane-terminated polymers
- Process for enantioselective enzymatic reduction of keto compounds
- RTV-1 silicone elastomer compositions which crosslink by means of alkoxy groups
Claims
1. A Hirudin derivative containing an N-terminal Ala-(SEQ ID NO: 8).
2. A thrombin inhibitor having a specific activity of at least 10,000 antithrombin units/mg containing an N-terminal Ala-(SEQ ID NO: 8).
3. A thrombin inhibitor having a specific activity in the range of 21,000 to 42,000 antithrombin units/ml containing an N-terminal Ala-(SEQ ID NO: 8).
4. The thrombin inhibitor according to claim 3, wherein said specific activity is 21,000 antithrombin units/ml.
5. The thrombin inhibitor according to claim 3, wherein said specific activity is 36,000 antithrombin units/ml.
6. The thrombin inhibitor according to claim 3, wherein said specific activity is 42,000 antithrombin units/ml.
| 4457866 | July 3, 1984 | Karges et al. |
| 4668662 | May 26, 1987 | Tripier |
| 5180668 | January 19, 1993 | Crause et al. |
| 4597785 | February 1986 | AUX |
| 5233686 | July 1986 | AUX |
| 5778786 | November 1986 | AUX |
| 7536687 | January 1988 | AUX |
| 4365585 | November 1989 | AUX |
| 0025190 | March 1981 | EPX |
| 158564 | October 1985 | EPX |
| 0168342 | January 1986 | EPX |
| 0171024 | February 1986 | EPX |
| 0200655 | November 1986 | EPX |
| 0225633 | June 1987 | EPX |
| 0252854 | January 1988 | EPX |
| 0338410 | October 1989 | EPX |
| 0352228 | January 1990 | EPX |
| 0356335 | February 1990 | EPX |
| 0161937 | October 1992 | EPX |
| 3445517 | June 1986 | DEX |
| 3900626 | July 1989 | DEX |
| 05143 | May 1990 | WOX |
- FEBS Letters, Bd. 202, Nr. 2, Jun. 1986, pp. 373-377; J. Dodt et al.: "Expression, secretion and processing of hirudin in E. coli using the alkaline phosphatase signal sequence". Applied Microbiology and Biotechnology, Bd. 34, No. 2, Nov. 1990, pp. 203-207, Springer-Verlag; E. Bender et al.: "Synthesis and secretion of hirudin by Streptomyces lividans". J.C. Janson Trends in Biotechnology vol. 2, No. 2, 1984. "Large Scale Affinity Purificaton-State of the Art and Future Prospects", pp. 31-38. "Hirudin: A Family of Iso-Proteins Isolation and Sequence Determination of New Hirudins," ISSN 0323-4347, Folia Haematol, Leipzig 115 (1988) 1-2, S. 30-35. "Primary Structures of New `Iso-hirudins`," FEBS Letters, vol. 225, No. 1, pp. 105-110 (1989). J.C. Janson Trends in Biotechnology vol. 2 (2) 1984. "Large Scale Affinity Purification" pp. 31-38.
Type: Grant
Filed: Nov 25, 1992
Date of Patent: Jul 6, 1999
Assignee: Consortium fur elektrochemische Industrie GmbH (Munich)
Inventors: Gerhard Schmid (Munich), Paul Habermann (Eppstein)
Primary Examiner: James Ketter
Law Firm: Collard & Roe, P.C.
Application Number: 7/982,064
International Classification: A61K 3858;
